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Anti Nucleobindin 2 / NEFA / Nesfatin precursor (NET1) polyclonal antibody


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Background

Nucleobindin 2 (NUCB2), also known as NEFA or Nesfatin precursor, is a ubiquitously expressed EF-hand Ca2+ binding protein that is implicated in various physiological processes. Nucleobindin 2 interacts with the postmitotic growth suppressor necdin in neurons. Both necdin and nucleobindin 2 are expressed in differentiated neurons and skeketal muscles and these proteins are likely to be involved in the regulation of survival and death of postmitotic cells by controlling Ca2+ homeostasis.

Data Link : Swiss-Prot P81117(mouse), Q9JI85(rat), P80303(human)

Application

1. Western blotting (1/3,000-1/1,000).
2. Immuno-precipitation.
3. Immuno-cytochemistry (dilution: 1/1,000)
4. Immuno-histochemistry (dilution: 1/2,000)
5. Immuno-electron microscopy
6. Immuno-affinity chromatography

Fig.1 Western blotting using this antibody.  Distribution of nucleobindin 2 and necdin in neonatal mouse organs. Homogenates of various organs from P0 mouse were separated by 10% SDS-PAGE and immunoblotted with this antibody (upper panel) and anti-necdin antibody (lower panel).  Nucleobindin 2 immunoreactive bands (~55 kDa) were detected in all the tissues examined. Tissue levels of nucleobindin 2 were high in the lung, brain, skeletal muscle, and spleen. On the other hand, necdin was enriched in the brain and skeletal muscle, and small amounts of necdin were detected in the kidney, lung, and heart.

Fi g.2 Immunocytochemistry for endogenous ucleobindin 2 and necdin in P19 cells. Cells were stained with this antibody and anti-necdin antibody by the avidin-biotin-peroxidase omplex method.  Left panel; undifferentiated P19 cells (UD). Right panels; enriched post-mitotic neurons (PN). Upper panels, nucleobindin 2 immuno-reactivity (Nucleobindin 2). Lower panels; necdin immuno eactivity (Necdin).  Nucleobindin 2 was localized to the cytoplasm near the nucleus in undifferentiated P19 cells, and its immunoreactivity in the cytoplasm was increased when P19 cells were induced to differentiate into neurons. On the other hand, the cytoplasm of undifferentiated cells was lightly stained for necdin, whose immunoreactivity was increased in both the cytoplasm and the nucleus of differentiated neurons.

Fig.3 Immunohistochemistry for nucleobindin 2 sing this antibody in neonatal mouse brain in vivo.  Frozen brain sections from P0 mouse were stained with this antibody by the avidin-biotin peroxidase complex method. A-C, cerebral cortex (parietal lobe). At higher magnification (B, C), fine granular immunoreactive materials are observed at both neuronal dendrites (arrows) and perikarya (asterisks) in the layer IV (arrowhead B in A) and subplate (arrowhead C in A) of the cerebral cortex. Scale bars, 100 um (A) and 10um (B and C).

<Reference>
This antibody was produced and used in ref.3.
1. Barnikol-Watanabe S et al (1994) “Human protein NEFA, a novel DNA binding /EF-hand/leucine zipper protein. Molecular cloning and sequence analysis of the cDNA, isolation and characterization of the protein.” Biol Chem Hoppe-Seyler 375: 497-512 PMID: 7811391
2. Kroll KA et al (1999) “Heterologous overexpression of human NEFA and studies on the two EF-hand calcium-binding sites.” Biochem Biophys Res Commun 260:1-8 PMID: 10381334
3. Taniguchi N et al (2000) “The postmitotic growth suppressor necdin interacts with a calcium-binding protein (NEFA) in neuronal cytoplasm.” J Biol Chem 275: 31674-31681 PMID: 10915798

Product List

Product Name Cat# Quantity Price

Anti Nucleobindin 2/ NEFA / Nesfatin precursor, Mouse (Rabbit)  DataSheet

BAM-73-109-EX

100 UL ¥34,500
$460

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.



“Anti Nucleobindin 2 / NEFA / Nesfatin precursor (NET1) polyclonal antibody” also belongs to the following categories.